Transactivation of the major capsid protein gene of herpes simplex virus type 1 requires a cellular transcription factor
| Autor: | R. L. Millette, L. K. Mills, S. Riddle, R. Wobig, R. Lown, P. Perry, Shin Chen |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
Transcriptional Activation
viruses Molecular Sequence Data Immunology Biology Binding Competitive Microbiology Immediate-Early Proteins Chloramphenicol acetyltransferase Viral Proteins Transactivation Capsid Virology Humans Simplexvirus Binding site Promoter Regions Genetic Transcription factor Gene Regulation of gene expression Binding Sites Base Sequence YY1 Promoter Molecular biology Insect Science DNA Viral Mutation Capsid Proteins Research Article HeLa Cells Transcription Factors |
| Zdroj: | Journal of Virology. 66:4304-4314 |
| ISSN: | 1098-5514 0022-538X |
| Popis: | The purpose of this investigation was to identify and characterize the regulatory elements involved in the transcriptional activation of the beta gamma (leaky-late or gamma 1) genes of herpes simplex virus type 1 (HSV-1) by using the major capsid protein (VP5 or ICP5) gene as model. Gel mobility shift assays with nuclear extracts from uninfected and infected HeLa cells enabled us to identify two major protein-DNA complexes involving the VP5 promoter. The mobilities of these two complexes remained unaltered, and no unique complexes were observed when infected cell nuclear extracts were used. DNase I and orthophenanthroline-Cu+ footprint analyses revealed that the two complexes involve a single binding site, GGCCATCTTGAA, located between -64 and -75 bp relative to the VP5 cap site. To determine the function of this leaky-late binding site (LBS) in VP5 gene activation, we tested the effect of mutations in this region by using transient expression of a cis-linked chloramphenicol acetyltransferase gene. Deletion of the above sequence resulted in a seven- to eightfold reduction in the level of transactivation of the chloramphenicol acetyltransferase gene by superinfection with HSV-1 or by cotransfection of HSV-1 immediate-early genes. From these results, we conclude that the LBS sequence and a cellular factor(s) are involved in the transactivation of the VP5 gene. A search of published gene sequences revealed that sequences related to the LBS exist in a number of other HSV-1, cytomegalovirus, retrovirus, and cellular promoters. Sequence homologies of binding sites and results of unpublished competition binding studies suggest that this leaky-late binding factor may be related to, or the same as, a ubiquitous cellular transcriptional factor called YY1 or common factor-1 (also known as NF-E1, delta, and UCRBP). |
| Databáze: | OpenAIRE |
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